Isolation and characterization of <i>n</i>-butane-utilizing microorganisms

McLee, A. G.; Kormendy, Agnes C.; Wayman, Morris

doi:10.1139/m72-186

Cited by 49 publications

(32 citation statements)

References 6 publications

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“…strain ATCC 27778. This strain was isolated from a naturaland domestic-gas-contaminated site and has been shown to utilize butane and butanol as growth substrates (10). Because most of the identified MTBE-cometabolizing bacteria utilize alkane as a feed substrate (6,8,17), we hypothesized that a culture that had been previously exposed to a mixture of hydrocarbons, including branched butane, would have the ability to produce the necessary enzymes for MTBE degradation.…”

Section: Resultsmentioning

confidence: 99%

Kinetics of Methyl t -Butyl Ether Cometabolism at Low Concentrations by Pure Cultures of Butane-Degrading Bacteria

Liu¹,

Speitel²,

Georgiou

2001

Appl Environ Microbiol

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Butane-oxidizing Arthrobacter (ATCC 27778) bacteria were shown to degrade low concentrations of methyl t-butyl ether (MTBE; range, 100 to 800 g/liter) with an apparent half-saturation concentration (K s ) of 2.14 mg/liter and a maximum substrate utilization rate (k c ) of 0.43 mg/mg of total suspended solids per day. Arthrobacter bacteria demonstrated MTBE degradation activity when grown on butane but not when grown on glucose, butanol, or tryptose phosphate broth. The presence of butane, tert-butyl alcohol, or acetylene had a negative impact on the MTBE degradation rate. Neither Methylosinus trichosporium OB3b nor Streptomyces griseus was able to cometabolize MTBE.The prevalent use of methyl t-butyl ether (MTBE) for gasoline oxygenation has led to its introduction into groundwater from spills and leaky underground storage tanks. MTBE is poorly adsorbed, chemically and biologically stable, and very soluble in water, making it very mobile and persistent in the environment. The U.S. Environmental Protection Agency has recently proposed scaling back the use of MTBE in gasoline in light of the increasing frequency with which MTBE is found as a groundwater contaminant nationwide (16; http://www.epa.gov /swerust1/mtbe/browner.pdf). Concentrations of MTBE in groundwater have been reported to range from 0.5 g/liter to Ͼ10 mg/liter. At least 20 states have established MTBE groundwater cleanup levels ranging from 20 to 400 g/liter for groundwater for potable use (http://www.epa.gov/OUST/mtbe /sumtable.htm). The U.S. Environmental Protection Agency has established a health advisory level of 20 to 40 g/liter for MTBE in drinking water (19).Efforts to develop bioremediation processes to help combat MTBE contamination of groundwater have been hampered by the recalcitrance of MTBE. The highly branched nature of MTBE resists most bacterial enzymatic attacks. Only a few pure and mixed bacterial cultures that are able to biodegrade MTBE have been identified (4,6,7,8,17).One potential MTBE biodegradation pathway involves the demethylation of MTBE to form tert-butyl alcohol (TBA) and formaldehyde (11, 17; K. L. Hurt, J. T. Wilson, and J. S. Cho, 5th Int. In Situ On-Site Bioremediation Symp., 1999), although the formation of tert-butyl formate from MTBE has also been observed (8). tert-Butyl formate is then hydrolyzed into TBA. In general, TBA is the first stable metabolite of MTBE, regardless of the type of bacterial cultures used. Hyman et al. (8) presented evidence that TBA is degraded by the same enzyme that degrades MTBE, although a soil microcosm was identified that was able to biodegrade TBA but not MTBE (M. J. Zenker, R. C. Borden, and M. A. Barlaz, 5th Int. In Situ On-Site Bioremediation Symp., 1999). TBA is biodegraded at a slower rate than MTBE (8, 17); thus, it tends to build up over time. If TBA is indeed degraded by the same enzyme that degrades MTBE, then the accumulation of TBA can have a detrimental effect on the MTBE biodegradation rate as a result of enzyme competition. Consequently, the effects of TBA on MTBE b...

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Section: Resultsmentioning

confidence: 99%

Kinetics of Methyl t -Butyl Ether Cometabolism at Low Concentrations by Pure Cultures of Butane-Degrading Bacteria

Liu¹,

Speitel²,

Georgiou

2001

Appl Environ Microbiol

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“…Growth on C 2 -C 4 gaseous alkanes is largely attributed to the Corynebacterium-Nocardia-Mycobacterium-Rhodococcus complex of Gram-positive bacteria (Ashraf et al, 1994;McLee et al, 1972;Perry, 1980). An exception to this association is the Gram-negative b-proteobacterium 'Pseudomonas butanovora' (ATCC 43655).…”

Section: Introductionmentioning

confidence: 99%

Butane monooxygenase of ‘Pseudomonas butanovora’: purification and biochemical characterization of a terminal-alkane hydroxylating diiron monooxygenase

2007

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Butane monooxygenase (sBMO) has been purified to homogeneity from the Gram-negative b-proteobacterium 'Pseudomonas butanovora' and confirmed to be a three-component diiron monooxygenase system. The reconstituted enzyme complex oxidized C 3 -C 6 linear and branched aliphatic alkanes, which are growth substrates for 'P. butanovora'. The sBMO complex was composed of an iron-containing hydroxylase (BMOH), a flavo-iron sulfur-containing NADH-oxidoreductase (BMOR) and a small regulatory component protein (BMOB). The physical characteristics of sBMO were remarkably similar to the sMMO family of soluble multicomponent diiron monooxgenases. However, the catalytic properties of sBMO were quantitatively different in regard to inactivation in the presence of substrate and product distribution. BMOH was capable of ethene oxidation when supplied with H 2 O 2 and ethene (known as the peroxide shunt), but this activity was at least three orders of magnitude less than that observed for the hydroxylase of sMMO of Methylosinus trichosporium OB3b. BMOH and BMOR were efficient in the oxidation of ethene in the absence of BMOB with regard to rate of reaction and product yield. Regiospecificity of sBMO was strongly biased towards primary hydroxylation, with ¢80 % of the hydroxylations occurring at the terminal carbon atom.

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“…Although a wide variety of microorganisms can utilize longchain, liquid alkanes, the ability to utilize short-chain, gaseous alkanes is mostly restricted to the Corynebacterium-NocardiaMycobacterium-Rhodococcus group of gram-positive bacteria (2,29). In addition, some gram-negative Pseudomonas spp.…”

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confidence: 99%

Two Distinct Monooxygenases for Alkane Oxidation in Nocardioides sp. Strain CF8

Hamamura¹,

Yeager²,

Arp

2001

Appl Environ Microbiol

110

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Alkane monooxygenases in Nocardioides sp. strain CF8 were examined at the physiological and genetic levels. Strain CF8 can utilize alkanes ranging in chain length from C 2 to C 16 . Butane degradation by butane-grown cells was strongly inhibited by allylthiourea, a copper-selective chelator, while hexane-, octane-, and decane-grown cells showed detectable butane degradation activity in the presence of allylthiourea. Growth on butane and hexane was strongly inhibited by 1-hexyne, while 1-hexyne did not affect growth on octane or decane. A specific 30-kDa acetylene-binding polypeptide was observed for butane-, hexane-, octane-, and decane-grown cells but was absent from cells grown with octane or decane in the presence of 1-hexyne. These results suggest the presence of two monooxygenases in strain CF8. Degenerate primers designed for PCR amplification of genes related to the binuclear-iron-containing alkane hydroxylase from Pseudomonas oleovorans were used to clone a related gene from strain CF8. Reverse transcription-PCR and Northern blot analysis showed that this gene encoding a binuclear-iron-containing alkane hydroxylase was expressed in cells grown on alkanes above C 6 . These results indicate the presence of two distinct monooxygenases for alkane oxidation in Nocardioides sp. strain CF8.

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Isolation and characterization of n-butane-utilizing microorganisms

Cited by 49 publications

References 6 publications

Kinetics of Methyl t -Butyl Ether Cometabolism at Low Concentrations by Pure Cultures of Butane-Degrading Bacteria

Kinetics of Methyl t -Butyl Ether Cometabolism at Low Concentrations by Pure Cultures of Butane-Degrading Bacteria

Butane monooxygenase of ‘Pseudomonas butanovora’: purification and biochemical characterization of a terminal-alkane hydroxylating diiron monooxygenase

Two Distinct Monooxygenases for Alkane Oxidation in Nocardioides sp. Strain CF8

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